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Using Bioluminescent Kinase Profiling Strips to Identify Kinase Inhibitor Selectivity and Promiscuity

  • Hicham ZegzoutiEmail author
  • Jacquelyn Hennek
  • Said A. GoueliEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1360)

Abstract

The advancement of a kinase inhibitor throughout drug discovery and development is predicated upon its selectivity towards the target of interest. Thus, profiling the compound against a broad panel of kinases is important for providing a better understanding of its activity and for obviating any off-target activities that can result in undesirable consequences. To assess the selectivity and potency of an inhibitor against multiple kinases, it is desirable to use a universal assay that can monitor the activity of all classes of kinases regardless of the nature of their substrates. The luminescent ADP-Glo kinase assay is a universal platform that measures kinase activity by quantifying the amount of the common kinase reaction product ADP. Here we present a method using standardized kinase profiling systems for inhibitor profiling studies based on ADP detection by luminescence. The kinase profiling systems are sets of kinases organized by family, presented in multi-tube strips containing eight enzymes, each with corresponding substrate strips, and standardized for optimal kinase activity. We show that using the kinase profiling strips we could quickly and easily generate multiple selectivity profiles using small or large kinase panels, and identify compound promiscuity within the kinome.

Key words

Kinase profiling Bioluminescence ADP detection Selectivity profiles Kinase assay 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Promega Corporation, Research and Development departmentMadisonUSA
  2. 2.University of Wisconsin School of Medicine and Public HealthMadisonUSA

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